Abstract
We previously identified a non-p53, p53-responsive DNA element (p53RE)-binding protein named NBP, functionally analogous to p53, from human cervical carcinoma Hela cells. Here we report a biochemical study demonstrating that this activity is the recently cloned p53 analog p63. NBP was purified through conventional and DNA affinity chromatography to apparent homogeneity with a prominent polypeptide migrating in between the 43 and 68 kDa positions on a SDS gel. This polypeptide immunoreacted with monoclonal anti-p63 but not anti-p53 or anti-p73 antibodies. Also, NBP co-purified with p63 through each step of fractionation, as detected with anti-p63 antibodies. DNA-protein complexes formed with purified NBP and p53RE-containing oligomers derived from the p21(waf1) promoter were supershifted by anti-p63 but not anti-p53 antibodies. Thus, these results demonstrate that NBP is encoded by the p53 homolog p63 gene.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Blotting, Western
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DNA-Binding Proteins / chemistry
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DNA-Binding Proteins / isolation & purification
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DNA-Binding Proteins / metabolism*
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Electrophoresis, Agar Gel
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Electrophoresis, Polyacrylamide Gel
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Female
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Genes, Tumor Suppressor
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HeLa Cells / chemistry
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Humans
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Immunoblotting
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Membrane Proteins*
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Nuclear Proteins / chemistry
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Nuclear Proteins / isolation & purification
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Nuclear Proteins / metabolism*
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Ovarian Neoplasms / pathology
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Phosphoproteins / chemistry
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Phosphoproteins / metabolism*
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Trans-Activators*
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Transcription Factors
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Tumor Suppressor Protein p53 / chemistry
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Tumor Suppressor Protein p53 / metabolism*
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Tumor Suppressor Proteins
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Uterine Cervical Neoplasms / pathology
Substances
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CKAP4 protein, human
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DNA-Binding Proteins
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Membrane Proteins
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Nuclear Proteins
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Phosphoproteins
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Son protein, mouse
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TP63 protein, human
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Trans-Activators
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Transcription Factors
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Trp63 protein, mouse
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Tumor Suppressor Protein p53
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Tumor Suppressor Proteins